The No. 1 Gators (29-2) posted the highest field goal percentage against Kentucky this season, knocking down 60 percent of their shots.
Four Florida players scored in double-figures, led by senior forward Patric Young with 18 points. Casey Prather followed with 15, while Scottie Wilbekin and Dorian Finney-Smith added 13 and 11, respectivley.
No. 25 Kentucky (22-9, 12-6 SEC) cut a 21-point halftime deficit to six midway through the second half, but the Gators closed with a 31-18 run over the final 11:52.
"I thought we fought with some good stuff, but when it came down to a six-, eight-point game, we took just two gawd-awful throws. They weren't even shots. They were throws," UK head coach Calipari said.
"When there's eight minutes to go in a game, it's time to win. You don't take those kind of shots. You just don't."
Last month in the first meeting between the teams, Florida outscored Kentucky 31-14 over the final 11:12 at Rupp Arena.
Wilbekin was key in both games. After UK sliced the deficit to six points, the senior guard scored seven points in the next three minutes to help the Gators reclaim a double-digit advantage.
Freshman forward Julius Randle led the Wildcats with 16 points and 10 rebounds, his 18th double-double of the season. James Young, another UK freshman, added 14 points, but managed just a lone free throw in the second half.
The Wildcats went almost four and a half minutes down the stretch without a field goal as Florida was able to build the lead back to 21 with 19 seconds remaining.
Kentucky returns to action on Friday in the SEC Tournament in Atlanta. The Wildcats will be the No. 2 seed playing at 7 p.m. ET on Friday, but their first opponent has yet to be determined.
• Kentucky leads the all-time series with Florida 94-36, including a 33-24 mark in Gainesville.
• The Wildcats dropped to 6-14 all-time when playing against the AP No. 1 team. UK is 1-2 against top-ranked Florida teams.
• UK's 44-22 deficit with 2:47 remaining in the first half was the largest it has faced this season.
• Andrew Harrison recorded a career-high three steals.